Home > Publications database > Untersuchung der Struktur und Dynamik von Bacteriorhodopsin-Intermediaten mit hochauflösenden Methoden |
Book/Report | FZJ-2020-00701 |
2001
Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag
Jülich
Please use a persistent id in citations: http://hdl.handle.net/2128/24116
Report No.: Juel-3845
Abstract: After excitation with a light quantum, the membrane protein bacteriorhodopsin starts aproton pumping reaction that can be divided into different intermediates. Structuralchanges of the protein during the pumping procedure play an important role. To solvethe mechanism of the pumping process it is important to analyse the structural changesor the structures of the intermediates, respectively. To determine the intermediatestructures with the x-ray method it is necessary to accumulate and capture the respectiveintermediate. In this work there are methods introduced and optimized to capture theintermediates of bacteriorhodopsin. A microspectrometer was developed to characterizeintermediates which were captured in protein crystals. This instrument allows themeasurements of infrared and visible absorption spectra simultanously. Procedures arepresented to capture the intermediated K, L and M in wild-type bacteriorhodopsin. Moreintermediates can be caught in bacteriorhodopsin mutants. A spectrometer for timeresolved measurements was built up to the characterize the photocycles of the mutants.The time resolved spectra of the mutants yield new insight to the proton pumpingreaction and they show the suitability for capturing intermediates. Finally the structureof the M-intermediate, captured with the described method and determined with the xraytechique, is shown and the findings .from this structure are described.
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